TIMELINE FOR RESEARCH

NOTE - see bottom of page for previous weeks' schedules and things to catch up on.
Definitions: 'cubby' = VDS cabinet in back of lab where all of our reagents are; 'pigeon hole' = the shelf for your lab notebook over by the window. Daily Schedule Summ14 Timeline F13

Looking for the Presentation Schedule? - see the Schedule Fall14 ( Semester)
  • This also has the Presentation Guidelines for all 3 types of presentations: Research Presentations, Technical Presentations, Journal Club Presentations
Link to Research Progress Table
Wikispaces check on every other Sunday (Bi-weekly)
Class slides are uploaded to CANVAS after each class.




This Week

This Week

WEEK15 - final week


Peer Reviews - should already have these done
  • Peer Review of Research Report on Target (Mid semester) - 1 per person
  • Peer Review of M&M Cloning - 2 per person
  • Peer Review of M&M Virtual- 2 per person
  • SKIP: peer review of M&M Expr, Pur, Char

Wet Lab work
finish cloning

express a FtHap protein if finished with at least 3 cloning attempts

Virtual Work
  • - analyze scores
    • ---InHouseCompounds (~40 ligs) -use 1 processor
    • ---CB306 (306 ligs) - use 1 processor
    • ---NIHClinicalCollection (446 ligs), use 1 processor
    • ---HF9PlatesPlates5_9.sdf (400 ligs) - use 1 processor

1-Dec Monday
2-Dec Tuesday
3-Dec Wednesday
class
FRI Survey (10-20 min)
https://utexas.qualtrics.com/SE/?SID=SV_6G8NU0TAFlSWawJ

4-Dec Thursday
Complete your 30 min of lab cleanup either today or tomorrow - see sheet on whiteboard
5-Dec Friday
Complete your 30 min of lab cleanup either today or tomorrow - see sheet on whiteboard
6-Dec Saturday
Final Lab Report Due @ 9pm

7-Dec Sunday

Wikispaces posting due - Sundays at 9 pm (For Weeks 14, 15)
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y'

Monday Dec 8th -
Final Lab Notebook Due at 9pm
WEEK15 - final week


Peer Reviews - should already have these done
  • Peer Review of Research Report on Target (Mid semester) - 1 per person
  • Peer Review of M&M Cloning - 2 per person
  • Peer Review of M&M Virtual- 2 per person
  • SKIP: peer review of M&M Expr, Pur, Char

Enzyme Assays & Inhibition Assays
  • determine if your enzyme is active
  • determine how much enzyme to use to get 'good' absorbance signal
  • vary amount of substrates(s) to optimize assay
  • Dilute inhibitors in DMSO
  • test out inhibitors (vary concentration)


Virtual Work
  • - analyze scores


1-Dec Monday
2-Dec Tuesday
3-Dec Wednesday
class
FRI Survey (10-20 min)
https://utexas.qualtrics.com/SE/?SID=SV_6G8NU0TAFlSWawJ

4-Dec Thursday
Complete your 30 min of lab cleanup either today or tomorrow - see sheet on whiteboard
5-Dec Friday
Complete your 30 min of lab cleanup either today or tomorrow - see sheet on whiteboard
6-Dec Saturday
Final Lab Report Due @ 9pm

7-Dec Sunday

Wikispaces posting due - Sundays at 9 pm (For Weeks 14, 15)
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y'

Monday Dec 8th -
Final Lab Notebook Due at 9pm
SPRINGERS
VETERANS




Next Week

Next Week





RECURRING ITEMS:
Wikispaces posting due - Sundays at 9 pm
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y'
Below that add your 'result' image for the week with brief commentary
This page will run in reverse chronological order - i.e. Newest stuff on top
Wiki postings should be a sort of highlights of results page.
You should have nanodrop images, gel images, yields from protein purification, brief virtual results, etc.
You don't need alot of lengthy description of the procedures - only figure captions and a brief analysis of
the result (e.g. 1-2 sentences of why it did or didn't work, next steps, etc.).
You should have enough information in the captiont to let the reader (Dr. B) understand what it is.

Lab Notebook check - Saturdays at 5pm

REQUIRED ITEMS FOR NOTEBOOKS:
All practice cloning work (including well labelled gels)
Protein work (esp protein gel and FPLC output and nanodrop)
We definitely need all of your Target work to be present in the notebook.
  • DNA Works Output file - print the text file and past in notebook
  • Tail Primer Design (protocol and what you designed)
  • Info from your Target Page (which means you need a Target page first!)

Update Target page on Wikispaces (need all the info from the list of items)
- if you don't have a Target page - start making it for your Target .


ITEMS FOR LATER:
Cut pNIC-Bsa4 that you grew up with BsaI (part of pNIC-Bsa4 Cloning protocol), gel check

Tail Primer Design and Ordering

Make LB+Kan plates (4) -
ProtocolLBAgarPlatesVDS.doc (not the Kan/Suc ones)

Make LB + Kan/Suc plates (6)
ProtocolLBAgarPlatesVDS.doc (not the Kan/Suc ones)






After Fire Safety Training is uploaded:
PAPER COPY – Print out of TxClass Safety Training -
- (can put in Safety section of VDS sign-in binder in lab)


Project Tasks (in rough order):
Nanodrop & Submit to DNA Sequencing (1.5 hrs)
Target Selection Form

Dilute some primers from lyophilized powder: random ones (pLIC-for & pLIC-rev)?
-- make working dilution in water from 100uM stock.
-- Store working dilution in YOUR own box.

Make 500ml of LB and autoclave (with mentor)
Transform pNIC-Bsa4 into DH5alpha (with a partner) - use 'homemade' competent cells (NOT THE NEB ones)
---- ProtocolTransformationforPlasmidPrep_pNIC_Bsa4_VDS_v5

Midi-prep (in pairs) - use Tony's new protocol
Input information about which culture of plasmid you have (or will) grow up.
GoogleDocs/Misc/Plasmid Concentrations
https://docs.google.com/spreadsheet/ccc?key=0AoO2KqKh2q_-dENvNGJEdW9qdUxQZmFYeTVoeXA0SUE&usp=sharing
Submit to DNA Sequencing for YOUR pNIC-Bsa4 that you grew up


Analyze DNA Sequence Exercise (as a group)

Design Overlap Primers ProtocolPCR_PrimerDesign_OverlapAssemblyVDS_Fall14.doc - 2 hrs
  • upload final primers to Google Docs/VDS/Misc/Primers/PrimerPlateOrderFall2014
  • post final primers to Wikispaces Target page


'My Second PCR' - PCR pLIC using pNIC-Bsa4
Set up target protein for Virtual Screening



Design Tail Primers for targets, order tail primers
Primer Dilution or your own primers

Create or Revise your Target page (not same as you Research Page)
LICpNIC-Bsa4cloningExerciseVDS_Fall14

Make Oligo mix for Targets

Expression
Purification
Characterization
PROTEIN INFO
Enter your protein yields into: Protein Spectrophotometer Calc
https://docs.google.com/spreadsheet/ccc?key=0AvGl3lMyhDsodC1mRVpNLVlDOUE4NlNxdldqNEZwRmc&usp=sharing

Virtual Work
CB Diversity (50K ligs) screening Due
M&M's
Cloning Materials & Methods due @ ???
-- see guidelines on GDocs

Peer Review of M&M Cloning is due @ ???
- you will have 2 to review
- make 5 constructive criticisms
- make 2 compliments

Virtual M&M due @ ???
Expression, Purification, Characterization M&M due @ ???
-- see guidelines on GDocs

Peer Review due ??:
M&M Virtual

Peer Review due ??:
M&M Expression, Pur, Char

Finish virtual screening
Enzyme Assay protocol
Inhibition Assay protocol


Graded Assignments:
HOURS:
Week1&2 Hrs
Week 3&4 Hrs
......
LabNotebook A, B, C, D

Wikispaces (Bi-Weekly)
WikiWeek1&2

Safety
Target Page
LICpNIC-Bsa4cloningExercise
Journal Club presentation and Question sheets
Technical Presentation (if applicable)
Research Presentation
PyMol Refresher
Virtual Screening Refresher
Mid-Semester Research Report on Target
Draft of M&M Cloning
Draft of M&M Virtual
Draft of M&M Express,Pur,Char
Final Report

2 Weeks Blocks layout

Week1
25-Aug
Monday
29-Aug
Friday
Week2
1-Sep
Monday
5-Sep
Friday
Week3
8-Sep
Monday
12-Sep
Friday
Week4
15-Sep
Monday
19-Sep
Friday
Week5
22-Sep
Monday
26-Sep
Friday
Week6
29-Sep
Monday
3-Oct
Friday
Week7
6-Oct
Monday
10-Oct
Friday
Week8
13-Oct
Monday
17-Oct
Friday
Week9
20-Oct
Monday
24-Oct
Friday
Week10
27-Oct
Monday
31-Oct
Friday
Week11
3-Nov
Monday
7-Nov
Friday
Week12
10-Nov
Monday
14-Nov
Friday
Week13
17-Nov
Monday
21-Nov
Friday
Week14
24-Nov
Tuesday
28-Nov
Friday
Week15
1-Dec
Monday
5-Dec
Friday




































WEEK1
WEEK2
WEEK3
WEEK4
WEEK5
WEEK6
WEEK7
WEEK8
WEEK9
WEEK10
WEEK11
WEEK12
WEEK13
WEEK14
WEEK15
8-Dec Monday
9-Dec Tuesday
10-DecWednesday
11-DecThursday
12-DecFriday
13-DecSaturday
14-DecSunday
15-DecMonday
16-DecTuesday
8-Sep Monday
9-Sep Tuesday
10-SepWednesday
11-SepThursday
12-SepFriday
13-SepSaturday
14-SepSunday
15-SepMonday
16-SepTuesday
17-SepWednesday
18-SepThursday
19-SepFriday
20-SepSaturday
21-SepSunday
22-SepMonday
23-SepTuesday
24-SepWednesday
25-SepThursday
26-SepFriday
27-SepSaturday
28-SepSunday
29-SepMonday
30-SepTuesday
1-Oct Wednesday
2-Oct Thursday
3-Oct Friday
4-Oct Saturday
5-Oct Sunday
6-Oct Monday
7-Oct Tuesday
8-Oct Wednesday
9-Oct Thursday
10-OctFriday
11-OctSaturday
12-OctSunday
13-OctMonday
14-OctTuesday
15-OctWednesday
16-OctThursday
17-OctFriday
18-OctSaturday
19-OctSunday
20-OctMonday
21-OctTuesday
22-OctWednesday
23-OctThursday
24-OctFriday
25-OctSaturday
26-OctSunday
27-OctMonday
28-OctTuesday
29-OctWednesday
30-OctThursday
31-OctFriday
1-Nov Saturday
2-Nov Sunday
3-Nov Monday
4-Nov Tuesday
5-Nov Wednesday
6-Nov Thursday
7-Nov Friday
8-Nov Saturday
9-Nov Sunday
10-Nov Monday
11-Nov Tuesday
12-Nov Wednesday
13-Nov Thursday
14-Nov Friday
15-Nov Saturday
16-Nov Sunday
17-Nov Monday
18-Nov Tuesday
19-Nov Wednesday
20-Nov Thursday
21-Nov Friday
22-Nov Saturday
23-Nov Sunday
24-Nov Monday
25-Nov Tuesday
26-Nov Wednesday
27-Nov Thursday
28-Nov Friday
29-Nov Saturday
30-Nov Sunday
1-Dec Monday
2-Dec Tuesday
3-Dec Wednesday
4-Dec Thursday
5-Dec Friday
6-Dec Saturday
7-Dec Sunday
8-Dec Monday
9-Dec Tuesday
10-DecWednesday
11-DecThursday
12-DecFriday
13-DecSaturday
14-DecSunday
15-DecMonday
16-DecTuesday














Target meetings:

1st one completed:





Primer Ordering Instructions (date?)

Everyone designs their primer - but we will only order one person's set.
Be sure you use the correct DNA Works output file to get your codon optimized DNA sequence (not just the CDS).
-- If you are use a Summer plate - you need to get your DNA Works Output file from Misc/Primers/DNAWorks_OutputSummer14
-- If you are use a Fall plate - you need to get your DNA Works Output file from Misc/Primers/DNAWorks_OutputFall14



Save the Word doc of your primer design into your GDocs
Print Word doc and paste into your Lab Notebook
Crosscheck your sequence with your Teammates
Put the sequences of the Tail Primers on your team's target page.
-- Enter your name for the person whose primers we will order - once you have designed the primers
Put the info and sequence on the Target Spreadsheet

https://docs.google.com/spreadsheet/ccc?key=0AvGl3lMyhDsodDJlZDlneXMtb1JuREV0MUZQYXU4YkE#gid=0

Order your primers
-- go to the ICMB core site (see our Targets Spreadsheet for the link)
-- find the IDT link
-- enter our username: VDS (all caps) and password: same as the password to get on to the laptops buildingname+labroomnumber
--Products/DNA/RNASynthesis/CustomDNAOligos
--select 2 for the Multiple Entry
-- leave defaults for Scale and Purification
-- enter your 2 primers and the Prefixes.
-- save to Wishlist, Dr. B will order when we have several of them.
WEEK1
25-Aug Monday
26-Aug Tuesday
27-Aug Wednesday
class
28-Aug Thursday
sign up for Fire Safety training (see link to left 'Required Lab Safety')
Lab times open for Nanodrop and Submit to DNA sequencing

Link to create dnaLIMS Account: (*must make an account before submitting DNA to sequencing)
http://146.6.133.14/cgi-bin/dna/loginNn.cgi

--- sign up on sheet above

Nanodrop protocol
Submit to DNA Sequencing


29-Aug Friday
30-Aug Saturday
31-Aug Sunday
WEEK1
25-Aug Monday
26-Aug Tuesday
27-Aug Wednesday
class
28-Aug Thursday
sign up for Fire Safety training (see link to left 'Required Lab Safety')
startup your cloning work again
29-Aug Friday
30-Aug Saturday
31-Aug Sunday
SPRINGERSWEEK2
Be sure you have 8-10hrs/week
- for the 1st week of class that should be about 3-4hrs since the lab was only open for 2 days
For this week, you need about 6.5-8 hrs.


ITEMS TO DO THIS WEEK**
  • sign up for Fire Safety training (see link to left 'Required Lab Safety')
  • Create your own folder in Google Docs under the folder: /StudentFolders
  • Make 500ml of LB and autoclave (with mentor) - anytime mentor available
  • Transform pNIC-Bsa4 into DH5alpha (with a partner) - use 'homemade' competent cells (NOT THE NEB ones) - at specific times
    • -- ProtocolTransformationforPlasmidPrep_pNIC_Bsa4_VDS_v5.doc
  • Dilute some primers (if they come in) - anytime mentor available
  • Target Selection Form - once Dr. B sends the link
  • Analyze DNA Sequence Exercise (as a group) - at specific times (this week or next depending on Target Selections and Primer Design)
  • (maybe) RE Digest - at specific times


1-Sep Monday - LABOR DAY - no lab
2-Sep Tuesday

Transformation times at 3:30 PM (must start on time)
Make 500ml of LB and autoclave (with mentor)


3-Sep Wednesday
class

Transformation times at 3:30 PM (must start on time)
Make 500ml of LB and autoclave (with mentor)


4-Sep Thursday


Make 500ml of LB and autoclave (with mentor)
Dilute some primers (if they come in)
Transformation times at 5:30 PM (must start on time)


5-Sep Friday

1:30-4:30 pm Analyze DNA Sequence Exercise (as a group) OR Primer Design if Targets figured out
Make 500ml of LB and autoclave (with mentor)
Dilute some primers (if they come in)


6-Sep Saturday
Lab Notebook check - Saturday at 5pm

What Goes Where for notebook?:
Nanorop - 2 images for Wiki & protocol/images in Notebook
Submit to DNA - protocol in Notebook, if have result - put in Notebook too
Wet lab and virtual work should go into notebook
Oligo Design - upload output file to GDocs, paste Oligo sequence into your Target page, in Notebook too
Analyze DNA Sequence - upload results to GDocs (put the protocol in your Notebook - results can stay in GDocs)
RE Digest - if done
PCR & run on an agarose gel - images to Wiki & protocol/images in Notebook


7-Sep Sunday
Target page updates due - 9 pm - SKIP FOR THIS WEEK

Wikispaces posting due - 9 pm
Add content to YOUR research page on Wikispaces
Put a line and 'Week 1&2'
Below that add your 'result' image for the week with brief commentary
This page will run in reverse chronological order - i.e. Newest stuff on top
Wiki postings should be a sort of highlights of results page.
You should have nanodrop images, gel images, yields from protein purification, brief virtual results, etc.
You don't need alot of lengthy description of the procedures - only figure captions and a brief analysis of
the result (e.g. 1-2 sentences of why it did or didn't work, next steps, etc.).
You should have enough information in the captiont to let the reader (Dr. B) understand what it is.
WEEK2
Be sure you have 8-10hrs/week
- for the 1st week of class that should be about 3-4hrs since the lab was only open for 2 days
For this week, you need about 6.5-8 hrs.

1-Sep Monday
2-Sep Tuesday
3-Sep Wednesday
class

3-4 Zain & Hailey site specific training with Priya

4-Sep Thursday
5-Sep Friday


6-Sep Saturday
7-Sep Sunday
WEEK3


ITEMS TO DO THIS WEEK
  • RE Digest of pGBR22
    • ProtocolRestrictionEnzymeDigestVDS.doc
  • Dilute some primers (if they come in) - anytime mentor available
  • Grow up your pNIC-Bsa4 colonies (DH5alpha) overnight
  • If failed last week, Re-transform pNIC-Bsa4 into DH5alpha (with a partner)
    • - use 'homemade' competent cells (NOT THE NEB ones) - at specific times
  • Target Selection Form - once Dr. B sends the link - SENT
  • Primer Design and Ordering
  • Create/Update Target pages
  • MAYBE: 'My First PCR' - using pGBR22
    • - ProtocolPCRVDS.xls


Grow up DH5alpha cells for Plasmid Preps (just store pellets though - don't do MidiPrep yet):

4:00 - 5 PM - start overnight cultures of plasmids in DH5alpha for midi prepping later.
-Grow up pNIC-Bsa4 overnight in 160ml of LB + Kan
--pNIC-Bsa4 is a 'low copy' plasmid
--spin down next morning
(save in -20degC or proceed to Midi-Prep - 2-3hrs)

-- if you don't' have time to spin down in morning (~30-45 min), you can
temporarily store your growth flask in 4degC for a few hours til you can spin it in the afternoon.
Pellets can be frozen and stored til next day or so.


In order to make lots of plasmid that we will need for cloning:
-- Plasmid Midi-prep (in pairs) - use Tony's new protocol

8-Sep Monday
9-Sep Tuesday
FIRE SAFETY TRAINING for FRI Students only
-- Speed Way in front of SAC (just show up and sign in)

Fill Out: Target Selection Form
https://docs.google.com/spreadsheet/viewform?formkey=dHJITzhKOHVvMUhNU05TRE81NktjV0E6MA

10-SepWednesday

class

Target Selection Form

11-SepThursday

Create/Update Target page on Wikispaces for your Team's target
Design Overlap Primers ProtocolPCR_PrimerDesign_OverlapAssemblyVDS_Fall14.doc - 2 hrs
  • upload final primers to Google Docs/VDS/Misc/Primers/PrimerPlateOrderFall2014
  • post final primers to Wikispaces Target page

12-SepFriday

Create/Update Target page on Wikispaces for your Team's target
Design Overlap Primers ProtocolPCR_PrimerDesign_OverlapAssemblyVDS_Fall14.doc - 2 hrs
  • upload final primers to Google Docs/VDS/Misc/Primers/PrimerPlateOrderFall2014
  • post final primers to Wikispaces Target page


13-SepSaturday
14-SepSunday
WEEK3
8-Sep Monday
9-Sep Tuesday
FIRE SAFETY TRAINING for FRI Students only
-- Speed Way in front of SAC (just show up and sign in)

10-SepWednesday

class


11-SepThursday

4- 8 pm PyMol Refresher Help Session in Computer Lab

12-SepFriday
PyMol Refresher due for Vets (less Zain, Hailey)
can count as 2 hours of lab time

13-SepSaturday
14-SepSunday
WEEK4


ITEMS TO DO THIS WEEK
  • Run gel of RE Digest of pGBR22
    • Agarose Gel protocol
  • Grow up your pNIC-Bsa4 colonies (DH5alpha) overnight (see below for steps)
  • Target Page - update yours with your team (the person who comes first on the list of names is the point person and delegates the work)
  • 'My First PCR' - using pGBR22
    • - ProtocolPCRVDS.xls
  • PyMol Refresher


Grow up pNIC-Bsa4 in DH5alpha cells for Plasmid Preps

4:00 - 5 PM - start overnight cultures of plasmids in DH5alpha for midi prepping later.
-Grow up pNIC-Bsa4 overnight in 160ml of LB + Kan
--pNIC-Bsa4 is a 'low copy' plasmid
--spin down next morning (10-11 am) - make sure a mentor can be there!
(save in -20degC or proceed to Midi-Prep - 2-3hrs)

-- if you don't' have time to spin down in morning (~30-45 min), you can
temporarily store your growth flask in 4degC for a few hours til you can spin it in the afternoon.
Pellets can be frozen and stored til next day or so.


Not due yet, but Mid Semester Research Report Guidelines posted to:
GDocs/Results&Data/ResearchReport/
GuidelinesResearchReportonTarget_VDS_Fall14.doc

15-SepMonday
16-SepTuesday
17-SepWednesday
18-SepThursday
19-SepFriday
20-SepSaturday


Lab Notebook check - Saturday at 5pm

What Goes Where for notebook?:
Oligo Design - protocol and DNA Works output file
Analyze DNA Sequence
RE Digest
PCR & run on an agarose gel (if done)


21-SepSunday
Wikispaces posting due - 9 pm
Add content to YOUR research page on Wikispaces
Put a line and 'Week 3&4'
Below that add your 'result' image for the week with brief commentary
This page will run in reverse chronological order - i.e. Newest stuff on top
Wiki postings should be a sort of highlights of results page.
You should have nanodrop images, gel images, yields from protein purification, brief virtual results, etc.
You don't need alot of lengthy description of the procedures - only figure captions and a brief analysis of
the result (e.g. 1-2 sentences of why it did or didn't work, next steps, etc.).
You should have enough information in the caption to let the reader (Dr. B) understand what it is.
WEEK4


Wet Lab
  • continue cloning, expression, purification, storage
  • look up your substrates and how to order them
  • look up enzyme assay in other research papers


Virtual Work

start Virtual Screening Refresher - see GDocs/Protocols/VirtualProtocols

Not due yet, but Mid Semester Research Report Guidelines posted to:
GDocs/Results&Data/ResearchReport/
GuidelinesResearchReportonTarget_VDS_Fall14.doc


15-SepMonday
16-SepTuesday
17-SepWednesday
18-SepThursday

Virtual Refresher Help Session

19-SepFriday
Virtual Refresher Help Session

20-SepSaturday


Lab Notebook check - Saturday at 5pm


21-SepSunday

Wikispaces posting due - 9 pm
WEEK5

  • Repeat - 'My First PCR' - if failed
  • PyMol Refresher
  • Target Page - update yours with your team (the person who comes first on the list of names is the point person and delegates the work)
  • Make Oligo Mix for your targets (each person makes own)
    • ProtocolPCR_PrimerOverlapWithQ5Polymerase_VDS_Fall14.docx

This week or next (depends on your lab hours)
  • Primary PCR
    • ProtocolPCR_PrimerOverlapWithQ5Polymerase_VDS_Fall14.docx
  • Midi-Prep your spun down pNIC-Bsa4
    • -- at appointed times
    • In order to make lots of plasmid that we will need for cloning:
    • -- Plasmid Midi-prep (in pairs) - use Tony's new protocol
  • Design Tail Primers for targets, order tail primers


Not due yet, but Mid Semester Research Report Guidelines posted to:
GDocs/Results&Data/ResearchReport/
GuidelinesResearchReportonTarget_VDS_Fall14.doc


22-SepMonday
23-SepTuesday
24-SepWednesday
PyMol Refresher due for Springers (plus Zain and Hailey)
can count as 2 hours of lab time
25-SepThursday


26-SepFriday
27-SepSaturday
28-SepSunday

Target page updates due - 9 pm
WEEK5

Wet Lab
  • continue cloning, expression, purification, storage
  • look up your substrates and how to order them
  • look up enzyme assay in other research papers

Virtual Work
  • Begin Virtual Screening setup (see meeting times)
    • create homology model if needed for your target
    • ProtocolHomologyModelVDS_v?.doc
      Protocol_LigandPrep_VDS_v1_?.docx
      ProtocolVirtualScreenYOURTarget_VDS_??.doc
You don't have to do all of these protocols right away - but you should aim to have your Homology model created this week and have started on Control Ligands selection (the first part of the Virtual Screening Protocol).
-- see the sign up spreadsheet above for times

Target Page - update yours with your team (the person who comes first on the list of names is the point person and delegates the work)

Not due yet, but Mid Semester Research Report Guidelines posted to:
GDocs/Results&Data/ResearchReport/
GuidelinesResearchReportonTarget_VDS_Fall14.d


22-SepMonday
23-SepTuesday
24-SepWednesday
Virtual Screening Refresher due for Vets
can count as 2 hours of lab time
(Zain & Hailey) do PyMol Refresher

25-SepThursday
26-SepFriday
27-SepSaturday
28-SepSunday

Target page updates due - 9 pm
SIGN UP FOR SPRINGERS FOR 1st Lab Times:
LINK TO EDIT THE PAGE: Add your name to the time you want



WEEK6

Check the Mentor schedule to see when mentors will be present in lab to help you out.


Wet Lab
  • Design Tail Primers for targets, order tail primers
    • ProtocolPCR_PrimerDesignTails_for_pNIC-Bsa4CloningFall14.doc
  • Primary PCR trials
    • ProtocolPCR_PrimerOverlapWithQ5Polymerase_VDS_Fall14.docx
  • Midi-Prep your spun down pNIC-Bsa4
    • -- at appointed times (sign up on spreadsheet)
    • In order to make lots of plasmid that we will need for cloning:
    • -- Plasmid Midi-prep (in pairs) - use Tony's new protocol
  • Secondary PCR trials

Virtual Work
Virtual Screening Refresher - start looking at will be due next week


29-SepMonday
  • Design Tail Primers for targets, order tail primers (sign up for times)

30-SepTuesday
  • Design Tail Primers for targets, order tail primers

1-Oct Wednesday
2-Oct Thursday
3-Oct Friday
4-Oct Saturday


5-Oct Sunday
Mid Semester Research Report due (a.k.a. Report on your Target)
Guidelines posted to:
GDocs/Results&Data/ResearchReport/GuidelinesResearchReportonTarget_VDS_Fall14.doc
WEEK6
Cloning should be completed by end or September
Start Expression


Virtual Work
  • Control Ligands selection and LigPrep is due at end of week
    • Beginning of the ProtocolVirtualScreenYOURTarget_VDS_Fall14.doc
    • Protocol_LigandPrep_VDS_v1_6.docx


29-SepMonday
30-SepTuesday
1-Oct Wednesday
2-Oct Thursday
3-Oct Friday
4-Oct Saturday
Lab Notebook check - Saturdays at 5pm

5-Oct Sunday
Mid Semester Research Report due (a.k.a. Report on your Target)
Guidelines posted to:
GDocs/Results&Data/ResearchReport/GuidelinesResearchReportonTarget_VDS_Fall14.doc
WEEK7

Check the Mentor schedule to see when mentors
will be present in lab to help you out.

Wet Lab
  • Primary PCR trials
    • ProtocolPCR_PrimerOverlap
    WithQ5Polymerase_VDS_Fall14.docx
  • Primer Dilution of your own tail primers

  • Secondary PCR trials


Virtual Work
Virtual Screening Refresher - due in 2 weeks from now

6-Oct Monday
7-Oct Tuesday
8-Oct Wednesday
9-Oct Thursday
10-OctFriday
11-OctSaturday
Lab Notebook check - Saturdays at 5pm


12-OctSunday


Wikispaces posting due - Sundays at 9 pm (For Weeks 5,6,7)
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y&Z'
WEEK7

Virtual Work
  • Control Ligands selection and LigPrep is due at end of week
    • Beginning of the ProtocolVirtualScreenYOURTarget_VDS_Fall14.doc
    • Protocol_LigandPrep_VDS_v1_6.docx
  • Virtual Set up is due at end of week
- e.g homology model (if needed) and setting up protein in GOLD
        • Submit docking job of control ligands is due at end of week
        • ProtocolHomologyModelVDS_v5.doc

- analyze control ligands' scores

We will set up


6-Oct Monday
7-Oct Tuesday
8-Oct Wednesday
9-Oct Thursday
10-OctFriday
11-OctSaturday
Lab Notebook check - Saturdays at 5pm

12-OctSunday

Wikispaces posting due - Sundays at 9 pm (For Weeks 5,6,7)
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y&Z'
WEEK8

Check the Mentor schedule to see when mentors will be present in lab to help you out.
We will also have some dedicated virtual screening help sessions (see below)

Wet Lab
  • continue cloning work on your project
    • start cutting your pNIC-Bsa4 while working on your PCRs
Virtual Work
* Virtual Screening Refresher (can count as 4 hrs of wet lab time for the whole exercise)

13-OctMonday
5-7 pm Virtual Screening Help Session in wet lab
14-OctTuesday
15-OctWednesday
3-7 pm Virtual Screening Help Session in wet lab

16-OctThursday
4-7 pm Help Session for:
Virtual Screening & LICpNIC-Bsa4cloningExercise

17-OctFriday
2-6 PM Help Session for:
Virtual Screening & LICpNIC-Bsa4cloningExercise

18-OctSaturday
1- 4 PM Help Session for:
Virtual Screening & LICpNIC-Bsa4cloningExercise

Run1 of Virtual Screen Refresher due today
LICpNIC-Bsa4cloningExerciseVDS_Fall14 due today

19-OctSunday
WEEK8

Wet Lab
  • protein expressions
  • purchase your substrates with Dr. B
  • enzyme assays

Virtual Work
- analyze control ligands' scores
- submit jobs for small screening libraries (cb306, NIHcc)
---InHouseCompounds (~40 ligs) -use 1 processor
---CB306 (306 ligs) - use 1 processor
---NIHClinicalCollection (446 ligs), use 1 processor
---HF9PlatesPlates5_9.sdf (400 ligs) - use 1 processor

--- If those look good - submit CBDiversity (large library - ~50k)


13-OctMonday
14-OctTuesday
15-OctWednesday
16-OctThursday
17-OctFriday

18-OctSaturday


19-OctSunday
WEEK9

Check the Mentor schedule to see when mentors will be present in lab to help you out.
We will also have some dedicated virtual screening help sessions (see below)

Wet Lab
  • continue cloning work on your project
    • start cutting your pNIC-Bsa4 while working on your PCRs
    • once PCR Squared is done - carry out cloning protocol
      • ProtocolpNIC-Bsa4_cloningVDS_Fall14.doc
      • make sucrose + kan plates if there aren't any (get a mentors help)
Virtual Work

* Virtual Screening Refresher (can count as 4 hrs of wet lab time for the whole exercise)

20-OctMonday

21-OctTuesday
22-OctWednesday
23-OctThursday
24-OctFriday
25-OctSaturday
Virtual Screening Refresher - due

26-OctSunday

Peer Review of Mid Semester Research Report on target
--- see GDrive folder for instructions (
ResearchReport_Materials&Methods_Guidelines)
WEEK9

Wet Lab
  • purchase your substrates with Dr. B (if you have not already done so)
  • enzyme assays - does your enzyme work?


Virtual Work
- analyze scores
- submit jobs for a large screening library (CB Diversity - 50k)
  • only use ~ 6 processors so that everyone can run their jobs

20-OctMonday

21-OctTuesday
22-OctWednesday
23-OctThursday
24-OctFriday
25-OctSaturday
26-OctSunday

Peer Review of Mid Semester Research Report on target
--- see GDrive folder for instructions (
ResearchReport_Materials&Methods_Guidelines)
WEEK10

Check the Mentor schedule to see when mentors will be present in lab to help you out.
We will also have some dedicated virtual screening help sessions (see below)

Wet Lab
  • continue cloning work on your project
  • carry out cloning protocol
    • ProtocolpNIC-Bsa4_cloningVDS_Fall14.doc
    • make sucrose + kan plates if there aren't any (get a mentors help)

Virtual Work
  • Control Ligands selection and LigPrep
  • Virtual Set up
  • Submit docking job of control ligands


27-OctMonday
28-OctTuesday
29-OctWednesday - Journal Club
30-OctThursday
31-OctFriday
1-Nov Saturday
Lab Notebook Due at 5pm

2-Nov Sunday

Cloning Materials & Methods due


Wikispaces posting due - Sundays at 9 pm (For Weeks 8,9,10)
Add content to YOUR research page on Wikispaces
Put a line and 'Week 8, 9, and 10'
WEEK10

Enzyme Assays
Virtual Screening
- ChemBridge Diversity
Priority: Luis, Charina, Andee, Nikki, Andrei, Hailey


27-OctMonday
28-OctTuesday
29-OctWednesday - Journal Club
30-OctThursday
31-OctFriday
1-Nov Saturday
Lab Notebook Due at 5pm

2-Nov Sunday

Cloning Materials & Methods due

Wikispaces posting due - Sundays at 9 pm (For Weeks 8,9,10)
Add content to YOUR research page on Wikispaces
Put a line and 'Week 8, 9, and 10'
WEEK11

Check the Mentor schedule to see when mentors will be present in lab to help you out.
We will also have some dedicated virtual screening help sessions (see below)

Wet Lab
  • continue cloning work on your project
  • carry out cloning protocol
    • ProtocolpNIC-Bsa4_cloningVDS_Fall14.doc
    • make sucrose + kan plates if there aren't any (get a mentors help)

Virtual Work
  • Control Ligands selection
    • ProtocolVirtualScreenYOURTarget_VDS_??.doc (first part only for now = Control Ligand Selection)


3-Nov Monday
4-Nov Tuesday
5-Nov Wednesday
6-Nov Thursday
7-Nov Friday
8-Nov Saturday
9-Nov Sunday

Peer Review of M&M Cloning Due at m/n -
–You have 2 different people’s which you need to review
–Make 5 constructive criticisms
–Make 2 compliments
- make sure to add your Initials to each comment so we know who did what - JTB
WEEK11

Enzyme Assays


3-Nov Monday

4-Nov Tuesday
5-Nov Wednesday
6-Nov Thursday
7-Nov Friday
8-Nov Saturday
9-Nov Sunday

Peer Review of M&M Cloning Due at m/n
–You have 2 different people’s which you need to review
–Make 5 constructive criticisms
–Make 2 compliments
- make sure to add your Initials to each comment so we know who did what - JTB
WEEK12

Virtual Work
  • Control Ligands - LigPrep
    • Protocol_LigandPrep_VDS_v1.docx
  • Virtual Set up
    • ProtocolVirtualScreenYOURTarget_VDS.doc
    • create homology model if needed for your target
      • ProtocolHomologyModelVDS_v.doc
  • Submit docking job of control ligands

Virtual Work
  • - analyze control ligands' scores
  • - submit jobs for small screening libraries (cb306, NIHcc, HF9)
  • ---InHouseCompounds (~40 ligs) -use 1 processor
    • ---CB306 (306 ligs) - use 1 processor
    • ---NIHClinicalCollection (446 ligs), use 1 processor
    • ---HF9PlatesPlates5_9.sdf (400 ligs) - use 1 processor


10-NovMonday
11-NovTuesday
12-NovWednesday
no journal club this week
13-Nov Thursday
14-Nov Friday
15-NovSaturday
16-NovSunday
Virtual M&M due
Enzyme Assays
  • determine is your enzyme is active
  • determine how much enzyme to use to get 'good' absorbance signal
  • vary amount of substrates(s) to optimize assay


Virtual Work
  • - analyze control ligands' scores
  • - submit jobs for small screening libraries (cb306, NIHcc, HF9)
  • ---InHouseCompounds (~40 ligs) -use 1 processor
    • ---CB306 (306 ligs) - use 1 processor
    • ---NIHClinicalCollection (446 ligs), use 1 processor
    • ---HF9PlatesPlates5_9.sdf (400 ligs) - use 1 processor
  • --- If those look good - submit CBDiversity (large library - ~50k)

WEEK12
10-NovMonday
11-NovTuesday
12-NovWednesday
no journal club this week
13-Nov Thursday
14-Nov Friday
15-NovSaturday
16-NovSunday
Virtual M&M due
WEEK13

Virtual Work
  • - analyze scores
  • - continue screening small screening libraries (cb306, NIHcc, HF9)
  • ---InHouseCompounds (~40 ligs) -use 1 processor
    • ---CB306 (306 ligs) - use 1 processor
    • ---NIHClinicalCollection (446 ligs), use 1 processor
    • ---HF9PlatesPlates5_9.sdf (400 ligs) - use 1 processor

17-Nov Monday
18-Nov Tuesday
19-Nov Wednesday
Journal Club

20-Nov Thursday
21-Nov Friday
22-Nov Saturday
Lab Notebook Due at 5pm

23-Nov Sunday

Wikispaces posting due - Sundays at 9 pm (For Weeks 11,12,13)
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y&Z'
WEEK13

Enzyme Assays & Inhibition Assays
  • determine if your enzyme is active
  • determine how much enzyme to use to get 'good' absorbance signal
  • vary amount of substrates(s) to optimize assay
  • Dilute inhibitors in DMSO
  • test out inhibitors (vary concentration)


Virtual Work
  • - analyze scores for ChemBridge Diversity

17-Nov Monday
18-Nov Tuesday
19-Nov Wednesday
Journal Club

20-Nov Thursday
21-Nov Friday
22-Nov Saturday
Lab Notebook Due at 5pm

23-Nov Sunday

Wikispaces posting due - Sundays at 9 pm (For Weeks 11,12,13)
Add content to YOUR research page on Wikispaces
Put a line and 'Week X&Y&Z'
Virtual Work
do some virtual screening

Wet Lab work
we will have mentors on duty for Mon, Tues, Wed - so continue your cloning work if you are in town

WEEK14
24-Nov Monday


25-Nov Tuesday -
Peer Review of M&M Virtual Due at m/n
–You have 2 different people’s which you need to review
–Make 5 constructive criticisms
–Make 2 compliments
- make sure to add your Initials to each comment so we know who did what - JTB

Expression, Purification, Characterization M&M due (can also turn in on Sunday night (Nov 30th)
  • It may be part of your proposed methods – but it still must be detailed.
  • I want you to think about the process and be able to explain the concepts.
  • You have done a somewhat similar Expression, Purification & Characterization in spring

26-Nov Wednesday - NO CLASS (Thanksgiving Break)
We will have mentors on duty if you want to do lab work


27-Nov Thursday
28-Nov Friday
29-Nov Saturday
30-Nov Sunday
see above for Peer Review and M&M that are due
Virtual Work
do some virtual screening

Wet Lab work
we will have mentors on duty for Mon, Tues, Wed


WEEK14
24-Nov Monday


25-Nov Tuesday
Peer Review of M&M Virtual Due at m/n
You have 2 different people’s which you need to review
–Make 5 constructive criticisms
–Make 2 compliments
- make sure to add your Initials to each comment so we know who did what - JTB

Expression, Purification, Characterization M&M due
(can also turn in on Sunday night (Nov 30th)

26-Nov Wednesday - NO CLASS (Thanksgiving Break)
We will have mentors on duty if you want to do lab work


27-Nov Thursday
28-Nov Friday
29-Nov Saturday
30-Nov Sunday
see above for Peer Review and M&M that are due